Ce este melanomul si cum apar metastazele?

Conferinta Asociatiei Melanom Romania: Inovatia in Melanom 2022 Viata este un drept pentru TOTI Bucuresti, 24-25 iunie 2022

Ce este melanomul si cum apar metastazele ?

Monica Neagu 1,2 1

Dept Imunologie, Institutul National de Patologie „Victor Babes” Bucuresti

2

Dept Patologie, Spitalul Clinic Colentina, Bucuresti

Identitatea celulara a unui melanocit normal

NHEM2 cells

Identitatea celulara a unui melanocit normal Melanocit Normal – expresia Pax3, Sox10, endothelin3 (ED-3), receptor specific (Endrb), c-Kit, Mitf ; UVR activeaza keratinocitele, activarea p53 induce proopiomelanocortina (POMC), precursorul 𝛼MSH si ACTH, hormoni cu activitate pro-pigmentare (Cui et al., 2007). 𝛼MSH activeaza MC1R care creste MITF - marker pentru melanocyte; MITF regleaza genele biogenezei melanosomilor (Hoek et al., 2008), sinteza de melanina si traficul melanozomilor (Chiaverini et al., 2008), sustinand in ansamblu procesele de fotoprotectie (Walker et al., 2009). Keratinocitele elibereaza 𝛼MSH, si multi alti factori (endotelina, GM-CSF, leukemia inhibitory factor - LIF, basic fibroblast growth factor - bFGF, hepatocyte growth factor – HGF) care induc proliferarea melanocitelor (Hirobe, 2005). Melanocitele normale au mecanisme anti-apoptotice rezistand la destructia ADN-ului la expunerea UV. Dupa expunerea la UV, expresia MITF creste si expresia genelor anti-apoptotice creste (BCL2, BCL2A1, ML-IAP) si a genelor care vor repara ADN (Haqa et al., 2013).

Monica Neagu, Carolina Constantin, Basak Engin, Iulia Popescu, Snapshot – changing melanocyte identity in melanoma developing route, Journal of Cell Identity, vol. 1, p. 0033-0047, October, 20, 2020

Melanocite in nevii benigni

Circulating benign melanocytes

Melanocite in nevii benigni Melanocitele din nevii melanocitari au particularitati proteomice si genomice; Majoritatea nevilor congenitali melanocitari au mutatii NRASQ61K/R in timp ce majoritatea nevilor aparuti in timpul vietii au mutatia BRAFV600E (Dadras, 2011); Daca in melanocitele normale se induc experimental genele NRAS sau BRAF ele capata un fenotip senescent – melanocitele din nevi au cresterea celulara oprita chiar si zeci de ani (Dadras, 2011). Melanocitele din nevii Benigni care au mutatii NRASQ61K sau BRAFV600E rar progreseaza la melanom. Dar daca melanocitele din nevi vor avea cateva proteine reglatorii nefunctionale (lipsa p16INK4a, Pten sau 𝛽-catenina) incepe procesul de tumorigeneza si aparitia melanomului din nevii benigni. Modificarile genetice/epigenetice combinate cu factori extrinseci vor duce la aparitia melanomului (Damsky et al., 2015).

Conditii care favorizeaza aparitia melanomului (melanomagenesis)

Micromediu inflammator Metabolism Factori Neuroendocrini Monica Neagu, et al, Snapshot – changing melanocyte identity in melanoma developing route, Journal of Cell Identity, vol. 1, p. 0033-0047, October, 20, 2020

Conditiicarefavorizeazamelanomageneza

Micromediu inflamator Proportia de celule imune – pattern-ul inflamator al tumorii caracterizeaza echilibrul intre un raspuns local eficient antitumoral si un micromediu tumoral pro‑tumoral (Zurac et al., 2013; Neagu et al., 2019); Tumora isi creaza propriul micromediu (TME) stabilind o retea de imuno supresie favorizanta dezvoltarii tumorale (Neagu et al, 2015). In TME celule stromale inter-relationeaza cu celulele immune si cu celulele din sistemul vascular. Moleculele din TME sunt preponderant citokine, chemokine si factori de crestere care fac tumora mai “invizibila” celulelor immune efectoare (tumor immune escape); Doua mecanisme distincte de rezistenta la atacul celulelor imune: existenta celulelor immune supresoare si lipsa de celule imune active. Daca infiltratul de celule imune e guvernat de CD8+ Ts, limfocite B si macrofage M2 se creaza un mediu imuno-supresor (Neagu et al., 2019); subpopulatii de limfocite NK care infiltreaza melanomul induc in TME procesul “tranzitia epithelial-mezenchimala” (EMT) care induce un fenotip agresiv al tumorii ( Izvoranu et al, 2021, Neagu et al., 2019); O inflamatie cronica, producerea si acumularea pe termen lung a factorilor pro-inflamatori (e.g. citokine/chemokine) induc intai local, apoi sistemic un mediu immuno-supresor care favorizeaza progresia tumorala ( Neagu et al., 2013, Neagu et al., 2015). Nivelele circulatorii de IL-6, IL-8 arata statusul inflamator al organismului si sunt asociate unui melanom care se dezvolta. In modele animale (Surcel et al., 2017) si la pacienti (Ene et al., 2015) IL-6, IL-8 este semnificativ crescut in corelatie cu evolutia clinica a tumorii.

Conditii care favorizeaza melanomageneza Caracteristici metabolice ale melanomagenezei Dereglarile metabolice in tumorigeneza sunt cel mai recent subiect de studiu O celula normala si o celula tumorala au status metabolic diferit. Metabolismul celulei tumorale este dinamic, are nevoie de un flux metabolic crescut si factori nutritionali crescuti pentru ca trebuie sa sustina o proliferare celulara accelerata, migrarea celulara si supravietuirea in conditii hipoxice adaptandu-se la diferite alte medii tisulare cand metastazeaza (Neagu, 2020). Metabolismul celulei tumorale genereaza productie crescuta de lactat, NO, ROS si prostaglandine – creaza un mediu inflamator favorizand metastazarea si adaptarea la un nou mediu (Netea-Maier et al., 2018). Metabolismul anormal induce expresia unor proteine disfunctionale care duc la cicluri celulare dereglate, scaderea proceselor anti-apoptotice, lipsa mortii celulare, cresterea capacitatii migratorii, cresterea capacitatii de adaptare la alte tesuturi decat cele in care au fost generate celulele tumorale (Wang et al., 2018). In melanom, metabolismul glucozei este ciclul cel mai dereglat, aceasta dereglare induce mai departe dereglari la nivel proteic si genetic (Gentric et al., 2017). Mitocondria, componentul celular major implicat in generarea energiei celulare, este implicata in tumorigeneza datorita faptului ca celula tumorala are cai de fosforilare oxidativa redusa si cai anabolice crescute. Mitocondria controleaza ROS, mutatiile ADN, instabilitatea genomica, controleaza fenomenul de autofagie si rezistenta la stimuli pro-apoptotici (Masgras et al., 2017; Neagu et al., 2019).

Conditii care favorizeaza melanomageneza Factori Neuroendocrini care favorizeaza melanomageneza In piele exista o serie de factori neuroendocrini implicati in procese fizio (pato) logice majore (Caruntu et al., 2014).

Catecolaminele pot induce progresia melanomului si pot stimula proliferarea celulelor tumorale prin intermediul receptorilor beta-adrenergici (Caruntu et al., 2014). Celulele tumorale de melanom au receptori care raspund la adrenalina. Celulele din tumorile primare cutanate si uveale sunt sensibile la adrenalina, in timp ce celulele din metastaze mult mai putin (Janik et al, 2017). Am aratat ca la concentratii crescute de epinefrina si norepinefrina celulele de melanom prolifereaza intens (Caruntu et al, 2014). Mai mult aceasta proliferare poate fi impiedicata de inhibitori specifici (Surcel et al., 2018).

Leptina, un hormon implicat in metabolismul lipidic poate induce inflamatie si actiune protumorala in melanom (Neagu, 2020, Constantin, 2022).

Melanocyte journey Proteomics

Genomics

Normal melanocyte

Melanoma Benign Nevi

Interrogating epigenome toward personalized approach in cutaneous melanoma, Elena-Georgiana Dobre, et al, J. Pers. Med. 2021, 11, 901. https://doi.org/10.3390/ jpm11090901

Interrogating epigenome toward personalized approach in cutaneous melanoma, Elena-Georgiana Dobre, et al, J. Pers. Med. 2021, 11, 901. https://doi.org/10.3390/ jpm11090901

Metastazarea • Exista 2 tipuri de migrare a melanocitelor tumorale – prin vasele limfatice si/sau prin circulatia sanguina (via limfatice sau via capilare sanguine de neoangiogeneza).

• Celulele tumorale vor circula in final prin ductul toracic sistemic • Celulele vor adera la capilarele diverselor tesuturi si vor extravaza • In unele tesuturi mai putin prietenoase desi vor intra in tesut vor genera o reactive imuna si vor fi eliminate • In unele tesuturi prietenoase pentru ca vor scapa de atacul celulelor imune vor prolifera intai in micro-metastaze si apoi metastaze decelabile imagistic

Cum metastazeaza melanomul?  Situsuri frecvente de metastazare: piele (metastaze satelit, metastaze in tranzit), ganglioni limfatici, plaman, creier, ficat,

sistem osos, intestin; Situsuri rare de metastazare: inima, pancreas, supra-renale, splina, stomac, esofag, tiroida, rinichi, sistemul genito-urinar (Damsky, 2011). Recurenta melanomului – intervale de 10 – 20 ani intre tumora primara si metastazare; fenomenul de dormanta tumorala semnalat acum 50 de ani (Fisher & Fisher, 1959) Mecanisme ale dormantei tumorale:  celulele raman in faza G0/G1 a ciclului celular  limfocitele citotoxice CD8+ T opresc proliferarea celulelor tumorale controlul apoptozei prin Fas/FasL micromediul tisular – celule tumorale dormante de melanom pot fi reactivate generand o metastaza de elementele tisulare legate de imbatranire (Fane, 2022) Mecanisme ale dormantei angiogenice Lipsa neo-vaselor care tin micrometastaza dormanta. Mecanisme ale dormantei imunologice Prezenta celulelor immune care limiteaza atat nr. celule tumorale cat si proliferarea acestora

Janowska, 2022

Concluzii Caracteristici celula tumorala: • 2000 - 6 proprietati • 2011 – se adauga inca doua “reprogramarea metabolismului celular” si “rezistenta la atacul sistemului imun” • se mai adauga “inflamatia pro-tumorala” care induce “instabilitatea genomului si mutatiile,” • 2021 sunt adaugate “plasticitatea fenotipica,” “reprogramarile epigenetice,” “microbiomul Hanahan D, 2022

polimorfic,” si “celulele senescente”

Bibliografie • Interrogating epigenome toward personalized approach in cutaneous melanoma, Elena-Georgiana Dobre, Carolina Constantin, Marieta Costache, Monica Neagu, J. Pers. Med. 2021, 11, 901. https://doi.org/10.3390/ jpm11090901 • Multi-Omics-Driven Biomarkers for Precision Medicine in Cutaneous Melanoma, Elena Georgiana Dobre, Monica Neagu, Biol. Life Sci. Forum 2021, 7(1), 20; https://doi.org/10.3390/ECB2021-10280 • Neuroendocrine factors in melanoma pathogenesis, Cristian Scheau, Carmen Draghici, Mihaela Adriana Ilie, Mihai Lupu, Iulia Solomon, Mircea Tampa, Simona Roxana Georgescu, Ana Caruntu, Carolina Constantin, Monica Neagu, Constantin Caruntu, Cancers, 2021 13(9), 2277; https://doi.org/10.3390/cancers13092277 • Therapeutic potential of interleukin‑15 in cancer Gheorghita Isvoranu, Mihaela Surcel, Adriana Narcisa Munteanu, Ovidiu Gabriel Bratu, Florentina Ionita‑Radu, Monica Teodora Neagu, Marioara Chiritoiu‑Butnaru, Experimental And Therapeutic Medicine 22: 675, 2021, https://www.spandidos-publications.com/10.3892/etm.2021.10107 • Snapshot – changing melanocyte identity in melanoma developing route, Monica Neagu, Carolina Constantin, Basak Engin, Iulia Popescu, Journal of Cell Identity, vol.

1, p. 0033-0047, October, 20, 2020.

• Metabolic traits in cutaneous melanoma, Monica Neagu, Frontiers in Oncology, 10: 851, 2020 doi: 10.3389/fonc.2020.00851 • Proteomic Technology “Lens” for Epithelial-Mesenchymal Transition Process Identification in Oncology, Monica Neagu, Carolina Constantin, et al, Analytical Cellular Pathology (Amsterdam) 2019:1-17, 2019 Oct 29;2019:3565970. doi: • Protein microarray technology: Assisting personalized medicine in oncology, Monica Neagu, Marinela Bostan and Carolina Constantin, World Academy of Sciences Journal, 1: 113-124, 2019.

• Inflammation and metabolism in cancer cell – mitochondria key player, Monica Neagu, Carolina Constantin, et al, Front Oncol, 9:348, 2019 doi: 10.3389/fonc.2019.00348 • Proteoglycans and immunobiology of cancer - therapeutic implications, George N Tzanakakis, Monica Neagu, Aristidis M Tsatsakis, Dragana Nikitovic, Frontiers in Immunology, Volume 10 | Article 875 doi: 10.3389/fimmu.2019.00875 1.

Immune based therapy for melanoma, Robert Ancuceanu, Monica Neagu, Indian Journal of Medical Research, Volume 143 Number 2, 135-144, 2016.

07/17/2022

16

Bibliografie • Natural killer cells monitoring in cutaneous melanoma – new dynamic biomarker, Gheorghița Isvoranu, Mihaela Surcel, […] Monica Neagu, Cornel Ursaciuc, Oncol Lett, 17: 4197-4206, 2019. DOI: 10.3892/ol.2019.10069 • Updates in immune-based multiplex assays, Cristiana Tanase, Radu Albulescu & Monica Neagu, J Immunoassay Immunochem, 2019, DOI: 10.1080/15321819.2019.1565064 • Multiplex assay for multiomics advances in personalized-precision medicine, Maria-Linda Popa, Monica Neagu, [..] Cristiana Tanase, J Immunoassay Immunochem, 2019, Published online: 11 Jan 2019, https://doi.org/10.1080/15321819.2018.156294 • Inflammation – key process in skin tumorigenesis, Monica Neagu, et al, Oncol Lett, (2019) 17: 4068-4084. https://doi.org/10.3892/ol.2018.9735 • Protein Microarray Technology for Antibody Detection Associated to Human Pathology, Carolina Constantin, [..] Monica Neagu, Romanian Archives of Microbiology and Immunology, Vol. 77, Issue 3, pp. 236-244, July-September, 2018.

• Adrenergic Modulation of Melanoma Cells Proliferation, Mihaela Surcel, […], Monica Neagu, Farmacia, 2018, Vol. 66, 5, 820-825.

• Positioning Europe for the EPITRANSCRIPTOMICS challenge. Jantsch MF, [..], Neagu M, [..], Fray R. RNA Biol. 2018 May 9:1-3. doi: 10.1080/15476286.2018.1460996 • Immunohistochemical features of partial regression in cutaneous melanoma – E-cadherin key molecule, Carmen Dumitru, […] Monica Neagu, [..], Sabina Zurac, HVM, Volume 9 | Issue 4, 166-177, 2017 • Inflammatory cytokine pattern is sex-dependent in mouse cutaneous melanoma experimental model, Mihaela Surcel, Carolina Constantin, Constantin Caruntu, Sabina Zurac, Monica Neagu, Journal of Immunology Research, Volume 2017, Article ID 9212134, 10 pages, https://doi.org/10.1155/2017/9212134 • Innovative array-based assay for omics pattern in melanoma, Carmen Dumitru, […] Monica Neagu, J Immunoassay Immunochem, 2017, vol 38, No. 4, pages 343354, http://dx.doi.org/10.1080/15321819.2017.1340898

07/17/2022

17

Bibliografie • Surface-Enhanced Laser Desorption/Ionization Mass Spectrometry for Biomarker Discovery in Cutaneous Melanoma, Carolina Constantin, Amanda Bulman, Diane McCarthy, Monica Neagu, Current Proteomics, 14 (2): 100-111, 2017.

• The Role of Estrogens and Estrogen Receptors in Melanoma Development and Progression, Caruntu C, […] Neagu M, Acta Endo (Buc) 2016, 12 (2): 234-241, doi: 10.4183/aeb.2016.234.

• Variations in the expression of TIMP1, TIMP2 and TIMP3 in cutaneous melanoma with regression and their possible function as prognostic predictors, Sabina Zurac, Monica Neagu, et al, Oncology Lett, 11: 3354-3360, 2016.

• Immune based therapy for melanoma, Robert Ancuceanu, Monica Neagu, Indian Journal of Medical Research, Volume 143 Number 2, 135-144, 2016.

• Interleukin 8 - Major Player in Cutaneous Melanoma Metastasis Process, Amalia-Elena Anghel, [..] Monica Neagu, Roum Biotechnological Letters, Vol. 20, No. 6, 10911- 10920, 2015.

•

25-OH Vitamin D and Interleukin-8 Emerging Biomarkers in Cutaneous Melanoma Development and Progression, Corina-Daniela Ene, Monica Neagu and Ilinca Nicolae, Mediators of Inflammation, pages 1-8, Article ID 904876, 2015.

• The relationship between interleukin 8 and Ki67 in cutaneous malignant melanoma, Amalia-Elena Anghel, [..] Monica Neagu, HVM Bioflux, Volume 7(3) 149-154, 2015.

• Chemokines in the Melanoma Metastasis Biomarkers Portrait. Neagu M, Constantin C, Longo C. J Immunoassay Immunochem. 2015;36(6):559-66.

• Inflammation markers in cutaneous melanoma - edgy biomarkers for prognosis, Monica Neagu, et al, Discoveries, 2015, Jan-Mar, 3(1): e38114 • Catecholamines increase in vitro proliferation of murine B16F10 melanoma cells, C Caruntu, […] M Neagu, Acta Endocrinologica (Buc) 2014, 10(4), 545-558 • Whole body microwave irradiation for improved dacarbazine therapeutical action in cutaneous melanoma mouse model, Monica Neagu, et al, Radiology Research and Practice, Radiology Research and Practice, vol. 2013, Article ID 414816, 10 pages, 2013. doi:10.1155/2013/414816 • Immune parameters in prognosis and therapy monitoring of cutaneous melanoma patients - experience, role and limitations, Monica Neagu, et al, BioMed Research International, Volume 2013, 15 November 2013 Article ID 107940

07/17/2022

18

Bibliografie Hallmarks of Cancer: New Dimensions, Hanahan D, CANCER DISCOVERY, 2022, doi: 10.1158/2159-8290.CD-21-1059 Central role of p53 in the suntan response and pathologic hyperpigmentation. Cui R, Widlund HR, Feigeetal E Cell 128(5): 853–864, 2007 Microphthalmia associated transcription factor regulates RAB27A gene expression and controls melanosome transport. Chiaverini C, Beuret L, Flori E et al, Journal of Biological Chemistry 283(18): 12635–12642, 2008 Novel MITF targets identified using a two-step DNA microarray strategy. Hoek KS, Schlegel NC, Eichhoff OM et al Pigment Cell & Melanoma Research 21(6): 665–676. 2008 Role of keratinocyte-derived factors involved in regulating the proliferation and differentiation of mammalian epidermal melanocytes.

Hirobe T, Pigment Cell Research 18(1): 2–12. 2005 BCL2A1 is a lineage-specific anti apoptotic melanoma oncogene that confers resistance to BRAF inhibition. Haqa R, Yokoyamab S, Hawryluk E B et al. Proc Natl Acad Sci USA 110(11): 4321–4326, 2013 Murine neonatal melanocytes exhibit a heightened proliferative response to ultraviolet radiation and migrate to the epidermal basal layer.

Walker GJ, Kimlin MG, Hacker E et al., Journal of Investigative Dermatology 129(1): 184–193. 2009 Molecular diagnostics in Melanoma: current status and perspectives, Dadras SS, Archives of Pathology and Laboratory Medicine 135(7): 860–869. 2011 𝛽catenin signaling controls metastasis in braf-activated pten deficient melanomas Damsky WE, Curley DP, Santhanakrishnan M et al.

Cancer Cell 20(6): 741–754. 2011 Metabolic changes in tumor cells and tumor-associated macrophages: a mutual relationship. Netea-Maier RT, Smit JWA, Netea MG Cancer Lett. 413: 102–9. 2018, doi: 10.1016/j.canlet.2017.10.037

Bibliografie Inflammasome and Cancer. Wang Z, Wang N, Zheng Y, Wang S , Exp Suppl. 108:281–302. 2018, doi: 10.1007/978-3-319-89390-7_12 The chaperone TRAP1 as a modulator of the mitochondrial adaptations in cancer cells. Masgras I, Sanchez-Martin C, Colombo G, Rasola AFront Oncol. 7:58. 2017doi: 10.3389/fonc.2017.00058 Diversified β-2-adrenergic Receptor Expression and Action in Melanoma Cells. Janik ME, Szlęzak D, Surman M, et al Anticancer Res.37(6):3025-3033, 2017.

Heterogeneity in cancer metabolism: new concepts in an old field. Gentric G, Mieulet V, Mechta-Grigoriou F, Antioxid Redox Signal 26:462– 85. doi: 10.1089/ars.2016.6750, 2017 Decoding Melanoma Metastasis, William E. Damsky, Jr., Lara E. Rosenbaum, Marcus Bosenberg Cancers 2011, 3, 126-163; doi:10.3390/cancers3010126 Experimental evidence in support of the dormant tumor cell. Fisher, B.; Fisher, E.R. Science, 1959, 130, 918-919 Stromal changes in the aged lung induce an emergence from melanoma dormancy, Mitchell E Fane et al2022 Jun;606(7913):396-405.doi: 10.1038/s41586-022-04774-2. Epub 2022 Jun 1.

Basis of Dormancy and Awakening in Cutaneous Metastatic Melanoma, Janowska, A.; Iannone, M.; Fidanzi, C.; Romanelli, M.; Filippi, L.; Del Re, M.; Martins, M.; Dini, V. The Genetic. Cancers 2022, 14, 2104. https://doi.org/10.3390/ cancers14092104

PN-III-P4-PCE-2021-0549 - Portrayal of antigen presenting cells in cutaneous melanoma - innovative pillars for harnessing immunotherapy

Multumesc! e-mail: neagu.monica@gmail.com